An enzymatic method for microdetermination of aphidicolin: a promising anticancer drug.
We have developed a method, based on the in vitro inhibition of purified human DNA polymerase alpha, the major enzyme of DNA replication, which allows the rapid and accurate determination of pmol amounts of aphidicolin, a promising anticancer drug. The efficacy of this simple method was verified by the determination of aphidicolin in the liver, spleen, blood and urine of mice treated parenterically with the drug. Given its sensitivity and the avoidance of radioactive tracers, this enzymatic method is suitable for the determination of the drug in body fluids and tissue biopsies from living humans. It allows the detection and quantitation of aphidicolin in the presence of inactive metabolite(s) with very similar chemical structure(s) such as those generated by liver microsomal oxidases. The technique will also be useful to monitor the purification of the drug from cultures of Cephalosporium aphidicola.
D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000032:analysis*; D016590:Aphidicolin; D002417:Cattle; D004257:DNA Polymerase II / Q000037:antagonists & inhibitors; D004224:Diterpenes / Q000032:analysis*; D008099:Liver / Q000032:analysis; D051379:Mice; D008832:Microchemistry / Q000379:methods; D013154:Spleen / Q000032:analysis
D000903:Antibiotics, Antineoplastic; D004224:Diterpenes; D016590:Aphidicolin; D004257:DNA Polymerase II